Selective suppression of lymphomas by functional loss of Hsf1 in a p53-deficient mouse model for spontaneous tumors.

A hallmark in the pathogenesis of cancer is the increased expression of heat shock proteins (Hsps) and other molecular chaperones observed in many tumor types, which is considered to be an adaptive response to enhance tumor cell survival. Heat shock transcription factor 1 (Hsf1) is a major transactivator of Hsp induction and has been proposed to affect tumor initiation and progression, regulating expression of Hsps and other molecular targets. In this report, we provide direct in vivo evidence that Hsf1 plays a critical role in the evolution of spontaneous tumors arising in p53(-/-) mice. Thus, loss of Hsf1 function did not prolong tumor-free survival, but surprisingly altered the spectrum of tumors that arose in p53(-/-) mice. Tumor development is rapid in p53(-/-) mice, which predominantly (about 70%) succumb to lymphomas. In contrast, hsf1(-/-)p53(-/-) mice rarely develop lymphomas (<8%), but succumb to other tumor types including testicular carcinomas and soft tissue sarcomas. Our findings suggest that an increase in p53-independent apoptotic cell death in association with altered cytokine signaling and suppressed production of inflammatory factors in hsf1(-/-) mice may contribute to selective lymphoma suppression. In conclusion, the data presented here link the loss of Hsf1-dependent function to decreased susceptibility to spontaneous lymphomagenesis, which may have implications for cancer prevention and therapy.

Regulation of rat heat shock factor 2 expression during the early organogenic phase of embryogenesis.

A central step in the transcriptional regulation of heat shock protein (hsp) genes is the binding of the heat shock factor (HSF) to the upstream heat shock elements (HSEs). In vertebrates, HSF2 has been suggested to mediate the transcriptional regulation of hsp gene expression during development and differentiation. The expression levels of HSF2 were shown to vary widely among fully developed mouse organs. However, there exists limited information on the regulation of HSF2 expression during the inductive stage of organ formation in mammalian development. In this study, we have cloned the rat HSF2 cDNA and examined embryos for HSF2 expression from days 9.5 (E9.5) to 15.5 (E15.5) of gestation that correspond to the period when the major organ primordia are being actively established. We show that rat HSF2 has 94.6 and 96.3% identity to mouse HSF2 in nucleotide and amino acid sequences, respectively. By establishing a competitive RT-PCR, we show that about 503.6 pg of HSF2 mRNA were present per microgram of embryonic RNA in the primitive streak stage E9.5 embryos. The amounts of HSF2 mRNA then gradually decreased, resulting in an approximately 300-fold reduction in E15.5 embryos. The amounts of HSF2 mRNA in the embryos were found to be closely correlated with those of HSF2 protein and their HSE-binding activities. To our knowledge, this is the first detailed report on the structure and regulation of the rat HSF2 during the early organogenic period of mammalian embryogenesis.

Role of small heat shock protein HSP25 in radioresistance and glutathione-redox cycle.

Expression of heat shock proteins (HSPs) has been shown to protect mammalian cells exposed to a variety of stress stimuli. Among various HSPs, small HSPs from diverse species were shown to protect cells against oxidative stress. Here, we show that the overexpression of the mouse small hsp gene, hsp25, provides protection against ionizing radiation. Our results demonstrate that the radiation survival of the L929 cells stably transfected with hsp25 was enhanced compared with that of the parental or vector transfected control, L25#1 cells. Our results also demonstrate that the radiation-induced apoptosis was reduced in HSP25 overexpressors. A detailed analysis of glutathione composition of those clones that overexpressed HSP25 revealed the increases of the glutathione pool, which primarily resulted from the increase of reduced glutathione. Our data suggest that higher content of GSH in HSP25 overexpressors was because of a faster reduction of oxidized glutathione (GSSG) to GSH rather than an increased de novo synthesis of GSH. The activities of glutathione reductase (GRd) and glutathione peroxidase (GPx) were greater in HSP25 overexpressors but the activity of gamma-glutamylcysteine synthetase was similar between the transfectants and the control cells. Consistent with our view, a steady state ratio of the GSH/GSSG was greater in the transfectants in comparison with the control L25#1 cells. A difference in the relative ratio became more significant after exposure to the ionizing radiation. To our knowledge, this study provides the first experimental evidence in support of the hypothesis that small HSP plays a key role in radioresistance by modulating the metabolism of glutathione. Based on the results obtained from the current investigation, we propose that HSP25 helps facilitate the glutathione-redox cycle and therefore, enhances glutathione utilization and maintains the cellular glutathione pool in favor of the reduced states.